IL-6 signaling mediates the germinal center response, IgM production and nociceptive sensitization in male mice after tibia fracture

Wen-Wu Li; Yang Yang; Tian-Zhi Guo; Peyman Sahbaie; Xiao-You Shi; Qin Guang; Wade S Kingery; Leonore A Herzenberg; J David Clark

doi:10.1016/j.bbi.2021.02.015

IL-6 signaling mediates the germinal center response, IgM production and nociceptive sensitization in male mice after tibia fracture

Brain Behav Immun. 2021 May:94:148-158. doi: 10.1016/j.bbi.2021.02.015. Epub 2021 Feb 23.

Authors

Wen-Wu Li¹, Yang Yang², Tian-Zhi Guo³, Peyman Sahbaie⁴, Xiao-You Shi⁵, Qin Guang⁶, Wade S Kingery⁷, Leonore A Herzenberg⁸, J David Clark⁹

Affiliations

¹ Anesthesiology Service, Veterans Affairs Palo Alto Health Care System, 3801 Miranda Ave, Palo Alto, CA 94304, USA; Department of Anesthesiology, Stanford University School of Medicine, 300 Pasture Drive, Stanford, CA 94305, USA.
² Department of Genetics, Stanford University School of Medicine, 300 Pasture Drive, Stanford, CA 94305, USA. Electronic address: yang71@stanford.edu.
³ Palo Alto Veterans Institute for Research, 3801 Miranda Ave, Palo Alto, CA 94304, USA.
⁴ Anesthesiology Service, Veterans Affairs Palo Alto Health Care System, 3801 Miranda Ave, Palo Alto, CA 94304, USA; Palo Alto Veterans Institute for Research, 3801 Miranda Ave, Palo Alto, CA 94304, USA. Electronic address: psahbaie@stanford.edu.
⁵ Anesthesiology Service, Veterans Affairs Palo Alto Health Care System, 3801 Miranda Ave, Palo Alto, CA 94304, USA; Department of Anesthesiology, Stanford University School of Medicine, 300 Pasture Drive, Stanford, CA 94305, USA. Electronic address: xyshi@stanford.edu.
⁶ Department of Genetics, Stanford University School of Medicine, 300 Pasture Drive, Stanford, CA 94305, USA.
⁷ Palo Alto Veterans Institute for Research, 3801 Miranda Ave, Palo Alto, CA 94304, USA. Electronic address: wkingery@stanford.edu.
⁸ Department of Genetics, Stanford University School of Medicine, 300 Pasture Drive, Stanford, CA 94305, USA. Electronic address: leeherz@stanford.edu.
⁹ Anesthesiology Service, Veterans Affairs Palo Alto Health Care System, 3801 Miranda Ave, Palo Alto, CA 94304, USA; Department of Anesthesiology, Stanford University School of Medicine, 300 Pasture Drive, Stanford, CA 94305, USA. Electronic address: djclark@stanford.edu.

Abstract

Background: Up-regulated interleukin 6 (IL-6) signaling, immune system activation, and pronociceptive autoantibodies are characteristic of complex regional pain syndrome (CRPS). IL-6 is known to promote B cell differentiation, thus we hypothesized that IL-6 signaling plays a crucial role in the development of adaptive immune responses and nociceptive sensitization in a murine tibia fracture model of CRPS.

Methods: Mice deficient in IL-6 expression (IL-6^-/-) or B cell deficient (muMT) underwent tibia fracture and 3 weeks of cast immobilization or sham injury. The deposition of IgM in fractured limbs was followed using Western blotting, and passive serum transfer to muMT fracture mice was used to detect nociception-supporting autoantibodies. Lymph nodes were assessed for hypertrophy, IL-6 expression was measured using qPCR and ELISA, and germinal center formation was evaluated using FACS and immunohistochemistry. The therapeutic effects of exogenous neutralizing anti-IL-6 antibodies were also evaluated in the CRPS fracture model.

Results: Functional IL-6 signaling was required for the post fracture development of nociceptive sensitization, vascular changes, and IgM immune complex deposition in the skin of injured limbs. Passive transfer of sera from wild-type, but not IL-6^-/- fracture mice into muMT fracture mice caused enhanced allodynia and postural unweighting. IL-6^-/- fracture mice displayed reduced popliteal lymphadenopathy after fracture. Germinal center responses were detected in the popliteal lymph nodes of wild-type, but not in IL-6^-/- fracture mice. We observed that IL-6 expression was dramatically enhanced in popliteal lymph node tissue after fracture. Conversely, administration of anti-IL-6 antibodies reduced nociceptive and vascular changes after fracture and inhibited lymphadenopathy.

Conclusions: Collectively, these data support the hypothesis that IL-6 signaling in the fracture limb of mice is required for germinal center formation, IgM autoantibody production and nociceptive sensitization. Anti-IL-6 therapies might, therefore, reduce pain after limb fracture or in the setting of CRPS.

Keywords: Autoimmunity; Complex regional pain syndrome; Fracture; Germinal center; Interleukin 6; Pain.

Published by Elsevier Inc.

Publication types

Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Complex Regional Pain Syndromes*
Disease Models, Animal
Germinal Center
Immunoglobulin M
Male
Mice
Nociception*
Tibia

Substances

Immunoglobulin M

Abstract

Publication types

MeSH terms

Substances

Grants and funding